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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Effects of polyamines on the insulin binding in isolated rat adipocytes were studied. In addition, the concentration of polyamines in adipose tissue was determined, and their localization revealed by fluorescence cytochemistry and immunocytochemistry. Spermine (0.1-10 mM) dose-dependently enhanced the insulin receptor binding; at a concentration of 5-10 mM spermine the insulin binding was enhanced by 100% above control values. Spermidine had a weaker effect than spermine whereas putrescine had no effect on insulin binding. Competition curves with unlabelled insulin indicated that spermine increased the insulin binding capacity without significantly affecting the binding affinity. Fluorescence and immunocytochemistry on adipose tissue localized the polyamines spermidine and/or spermine almost exclusively to the thin layer of adipocyte cytoplasm surrounding the lipid droplet. In addition, chemical analysis for polyamines in the adipocyte medium and in the cells indicated that a differential release of polyamines may have occurred from the cells. The possibility that polyamines may act as intracellular or intercellular (autocrine) regulators, modulating insulin binding, is discussed.
Mol Cell Endocrinol 1989 Apr
PMID:Polyamines in rat adipocytes: their localization and their effects on the insulin receptor binding. 266 68

The specific binding of insulin to either intact or Triton-solubilized Daudi cells (a Burkitt lymphoma cell line) was reduced by over 95% compared to that to control IM-9 lymphocytes due to a decrease in receptor number without a change in affinity. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed that 125I-labeled Daudi cells had reduced amounts (approximately 1/20th) of immunoprecipitable binding (alpha) subunit [mol wt (Mr), 130,000] of the receptor and a relatively abundant 210,000 Mr form not seen in IM-9 cells. The transmembranous (beta) subunit (Mr, 90,000) of the receptor, although not detected by 125I surface labeling, could be phosphorylated and, together with the 210,000 Mr form, exhibited the same 2-fold stimulation of phosphorylation by insulin as that in IM-9 cells. Northern blot hybridization revealed a decrease in Daudi cells of all four major species of insulin receptor mRNA. The Raji cell, another Burkitt lymphoma cell line, also exhibited reduced protein and genetic expression of the insulin receptor, indicating that reduced insulin receptor expression may be representative of other Burkitt lymphoma cell lines.
Mol Endocrinol 1989 Mar
PMID:Insulin receptor expression in the Burkitt lymphoma cells Daudi and Raji. 266 77

Biological activity and interference with insulin receptor complex fate of two modified sequences of insulin B21-B26, beta-Ala-Arg-Gly-Phe-Phe-Tyr-NH2 (DP-432) and beta-Ala-Arg-Pro-Phe-Phe-Tyr-NH2 (DP-640), were studied in cultured 18-day-old fetal rat hepatocytes known to respond to insulin by an acute stimulation of glycogenesis. The two derivatives stimulated [14C]glucose incorporation into glycogen in the absence of insulin independently of the deprivation of serum in the medium. The maximal effect of 3 mM DP-640 after 2 h, more pronounced than with 3 mM DP-432, was of the same order as that obtained with 10 nM insulin alone (stimulation index: 4.7 +/- 0.7, 2.5 +/- 0.2 and 3.6 +/- 0.9, n = 4, with DP-640, DP-432 and insulin, respectively) whereas insulin B-chain decreased glycogen labeling. Simultaneous addition of derivatives and insulin at maximal concentrations produced nearly additive effects. DP-640, as well as DP-432, increased the amount of [125I](A14) or (B26) human insulin associated with cells at 37 degrees C and inhibited intracellular insulin degradation with differences depending on the kind of insulin isomer and derivative, while the rapid insulin receptor cycle was not affected. Thus, the two derivatives specifically modified the cellular processing of insulin in cultured fetal hepatocytes, and exerted an insulin-like effect on glycogenesis clearly enhanced through modification of DP-432 by substitution of glycine for proline (DP-640).
Mol Cell Endocrinol 1989 Oct
PMID:Effects of modified insulin B21-B26 fragments on glycogenesis and on insulin-receptor complex fate in cultured fetal hepatocytes. 269 57

Two previously reported insulin receptor cDNA sequences differ by 36 base pairs (bp) in the distal alpha-subunit, suggesting that alternative mRNA splicing within the coding region may occur (two insulin receptor isoforms). We developed a quantitative modification of the polymerase chain reaction technique in order to detect and characterize differential mRNA splicing at this site within the distal alpha-subunit. Using RNA derived from a variety of human cell types, we detected two polymerase chain reaction-amplified cDNA species reflecting the presence or absence of the above 36 nucleotides. Identity of the two cDNA species was confirmed by Southern blots, the use of a BANI restriction site present only in the 36 base pair segment and dideoxy sequencing. The relative expression of the two mRNA forms varied markedly in a tissue-specific manner. Buffy coat leukocytes and Epstein-Barr virus-transformed lymphocytes express only the shorter mRNA. Placenta expresses both species equally; muscle, isolated adipocytes and cultured fibroblasts express somewhat more of the longer mRNA (relative ratios of mRNA abundance of 1.51, 3.18, and 2.77, respectively); liver expresses mostly the longer mRNA (relative ratio of 9.8). In RNA derived from cultured and fresh cells from patients with several states of insulin resistance, the relative expression of the two mRNA species was similar to results obtained with comparable normal tissues. Although the functional significance of alternative splicing of the insulin receptor mRNA is unknown, differential expression of these two receptor mRNAs may provide a structural basis for previously observed tissue-specific differences in insulin binding and action.
Mol Endocrinol 1989 Aug
PMID:Tissue-specific expression of two alternatively spliced insulin receptor mRNAs in man. 277 82

The tyrosine kinase activity of the epidermal growth factor (EGF) receptor is regulated by a truncated receptor of 100 kilodaltons (kDa) that contains the EGF-binding site but not the kinase domain. The inhibition of kinase is not due to competition for available EGF or for the kinase substrate-binding site. Chemical cross-linking studies suggest that the 100-kDa receptor may form a heterodimer with the intact EGF receptor. Structurally related receptor kinases, such as the platelet-derived growth factor receptor, the insulin receptor, and the Neu receptor, were not inhibited by the 100-kDa receptor. The results indicate that (i) the inhibition was specific for the EGF receptor, (ii) the kinase domain had little or no role in determining target specificity, and (iii) the regulation of kinase may be due to a specific interaction of the 100-kDa receptor with the ligand-binding domain of the EGF receptor kinase.
Mol Cell Biol 1989 Feb
PMID:Inhibition of tyrosine kinase activity of the epidermal growth factor (EGF) receptor by a truncated receptor form that binds to EGF: role for interreceptor interaction in kinase regulation. 278 40

The effect of a polyclonal anti-insulin receptor antibody (pIgG) on the insulin receptor tyrosine kinase (IRTK) activity toward poly-(Glu-Tyr) was examined using wheat germ agglutinin agarose-purified insulin receptors from rat liver membranes. The main effect of pIgG was a reduction of Vmax (from 60.8 to 31.8 pmol/min/mg), without changes of Km, when IRTK was activated by insulin. In contrast, when IRTK was activated by ATP preincubation, pIgG was unable to affect the reaction, suggesting that IRTK possesses at least two regulatory mechanisms, one of which can be affected by pIgG.
Mol Cell Endocrinol 1987 Sep
PMID:Regulation of insulin receptor-associated tyrosine kinase by a polyclonal IgG. 282 10

Studies investigating the effects of beta-naphthoflavone (beta NF) on insulin receptor binding and its intrinsic protein kinase activity in rat liver and placenta were performed. Membranes were prepared from maternal liver and placenta on gestation day 11 and used for [125I]insulin radioreceptor assay. Scatchard analysis showed that association constants (Ka) for high affinity binding sites were similar for placental and liver membrane. The administration of beta NF, 15 mg/kg, 1 day before study did not alter the specific binding of insulin to liver membranes, whereas ligand binding to placental preparations was decreased 40% from control. Scatchard analysis of binding to placental membranes suggests that beta NF treatment was associated with a change in the number of high affinity binding sites. In further studies membrane receptors were solubilized and partially purified by wheat germ agglutinin affinity chromatography for protein kinase assay. Insulin stimulated the phosphorylation of the Mr 95,000 subunit of the receptor in lectin-purified membrane proteins from liver and placenta. In liver receptor preparations, beta NF treatment was associated with a nearly 3-fold increase in the insulin-stimulated phosphorylation of the 95-kD protein. In contrast, placental receptor preparations showed a 40% decrease in the extent of autophosphorylation following beta NF treatment. Insulin-stimulated phosphorylation of an exogenous substrate poly(Glu4, Tyr) also showed a divergent pattern of changes in liver and placental receptors following beta NF treatment. In studies during late gestation (day 18), beta NF treatment was also associated with an increase in liver receptor kinase activity, whereas placental receptors showed a decrease in autophosphorylation. Thus, acute treatment with beta NF during mid and late gestation was associated with significant alterations in insulin receptor protein kinase activity, and data suggest that fetal insulin receptors may respond in a different manner than maternal receptors to polyaromatic compounds like beta NF. The observed effects of beta NF on liver and placental receptor kinase activity may be related to alterations in insulin function in the regulation of pregnancy and fetoplacental growth.
Mol Pharmacol 1988 Mar
PMID:Effects of beta-naphthoflavone on insulin receptor binding and protein kinase activity in rat liver and placenta. 283 20

The insulin receptor plays a central role in mediating the biological actions of insulin. We have used Epstein-Barr virus-transformed lymphocytes (EBV-lymphocytes) to investigate the receptor defects in patients with genetic forms of insulin resistance. Within the normal population, we found a close correlation between the number of insulin receptors on the surface of EBV-lymphocytes and the cellular content of insulin receptor mRNA. In addition, we have used the cloned human insulin receptor cDNA to investigate the nature of the mutations causing the reduction in the number of insulin receptors in EBV-lymphocytes from three insulin resistant patients. One patient with leprechaunism has a marked reduction in the level of receptor mRNA, which probably accounts for the extremely slow rate of receptor biosynthesis measured in this patient's cells. The remaining two patients with type A extreme insulin resistance are sisters, the products of a consanguineous marriage, who have normal levels of insulin receptor mRNA. We have previously shown that the insulin receptor precursor is synthesized at a normal rate in these patients' cells, thus suggesting a defect in the posttranslational processing of the receptor or in its translocation to the plasma membrane.
Mol Endocrinol 1988 Mar
PMID:Defects in human insulin receptor gene expression. 284 May 73

Insulin stimulates the autophosphorylation of the beta-subunit of the insulin receptor (IR) on tyrosine residues. Mutations which compromise IR autophosphorylation in vivo result in a decrease of the insulin-activated uptake of 2-deoxyglucose. These results are consistent with previous results which implicate IR autophosphorylation in the generation of the insulin response by cells. To further explore the specificity of the IR tyrosine phosphokinase (TPK) domain in IR function, we have altered the human IR (hIR) cDNA to encode truncated insulin-independent TPKs, which are expressed in chinese hamster ovary (CHO) cells as either membrane-anchored or cytosolic proteins. Both mutant hIRs exhibit TPK activity in vitro, although the cytosolic form is approximately 20 times more active. The carbohydrate moiety of the membrane-anchored form is of the high mannose type, consistent with an intracellular localization for this mutant hIR. The two mutant hIRs mediate very different physiological responses in transfected cells: the membrane-anchored, but not the cytosolic, hIR TPK mediates a constitutively elevated (135% the maximum insulin-stimulated response in CHO cells) insulin-independent uptake of 2-deoxyglucose. These results thus suggest that the hIR TPK is in fact specific for this aspect of IR function and, when membrane-associated, can mediate the insulin-independent uptake of 2-deoxyglucose. Neither of these mutant hIRs appears to transform CHO cells.
Mol Endocrinol 1987 Jan
PMID:A membrane-anchored cytoplasmic domain of the human insulin receptor mediates a constitutively elevated insulin-independent uptake of 2-deoxyglucose. 284 59

Studies characterized insulin and EGF receptors in human placental tissue from smokers and nonsmokers. Specific binding of 125I-labeled insulin and EGF to placental membranes was not different for nonsmokers compared with smokers. EGF and insulin receptor kinases were further studied using a wheat germ agglutinin-purified preparation of solubilized placental membrane proteins. In extracts from the nonsmoker group, EGF stimulated the active phosphorylation of Mr 170,000 and 140,000 protein bands, which was half-maximal (EC50) at 5 x 10(-8) M. In extracts from the smokers group, however, phosphorylation of these two protein bands was barely detectable over a range of 0 to 10(-6) M EGF. Thus, EGF-stimulated phosphorylation of the 170,000 and 140,000 bands was markedly decreased in placental membranes from smokers. In contrast, insulin stimulated the phosphorylation of a 95,000 protein that was immunoprecipitated with anti-insulin receptor antiserum in membrane preparations from both nonsmokers and smokers. Dose-response curves for autophosphorylation indicate that EC50 values were 2.6 and 7.0 nM insulin for nonsmokers and smokers, respectively. Laser densitometry scan of the 95,000 band on autoradiograms further showed that maximal 32P incorporation was 30% greater in smokers compared with nonsmokers. Analysis of the insulin-dependent phosphorylation of an exogenous substrate, poly(Glu,Tyr) (4:1), showed a similar pattern of values for nonsmokers versus smokers. These results indicate that insulin receptor autophosphorylation and tyrosine kinase activity were normal or increased, whereas EGF-stimulated kinase activity was markedly decreased in placental membrane proteins from smokers. Western blot analysis using an antiserum to the EGF receptor showed the presence of immunoreactive bands of 126,000 and 150,000-170,000 in receptor preparations from nonsmokers, whereas only the 126,000 protein was detected in preparations from smokers. Thus, the smoking-related deficiency in EGF receptor autophosphorylation appeared to be due to the absence of a 150,000-170,000 receptor protein. In conclusion, maternal cigarette smoking is associated with selective alterations in two major receptor-mediated pathways thought to be involved in cell growth and differentiation in human placenta.
Mol Pharmacol 1988 Sep
PMID:Smoking-related alterations in epidermal growth factor and insulin receptors in human placenta. 284 46


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